Agilent Bio-Columns – Critical Quality Attributes application compendium


Introduction 05
1 Titer determination 07
1.1 Background 07
1.2 Getting Started 08
1.3 How to Guide – Affinity Chromatography for Titer Determination 09
1.4 Featured Application Notes 16
1.5 Cell Clone Selection Using the Agilent Bio-Monolith Protein A
Column and LC/MS. – 5991-5124EN 16
1.6 mAb Titer Analysis with the Agilent Bio-Monolith Protein A Column
– 5991-5135EN 24
1.7 Additional Application Notes 31
1.8 More Information 31
2 Intact & Subunit Purity 32
2.1 Background 32
2.2 Getting Started 33
2.3 Featured Application Notes 35
2.4 Fast and High Resolution Analysis of Intact and Reduced
Therapeutic Monoclonal Antibodies (mAbs) – 5991-6274EN 35
2.5 Disulfide Linkage Analysis of IgG1 using an Agilent 1260 Infinity
Bio-inert LC System with an Agilent ZORBAX RRHD Diphenyl
sub-2 μm Column – 5991-1694EN 40
2.6 Determination of Drug-to-Antibody Distribution in Cysteine-Linked
ADCs – 5991-7192EN 45
2.7 Additional Application Notes 49
3 Peptide mapping 50
3.1 Background 50
3.2 Getting Started 51
3.3 How to Guide – Peptide Mapping
Keys for Enabling Optimum Peptide Characterizations
– 5991-2348EN 52
3.4 Featured Application Notes 76
3.5 Reproducible LC/MS Peptide Separation Using Agilent AdvanceBio
Peptide Plus Columns – 5991-8067EN 76
3.6 High-Resolution Mapping of Drug Conjugated Peptides
in an ADC Digest- 5991-6191EN 80
3.7 Analysis of Tryptic Digests of a Monoclonal Antibody
and an Antibody-Drug Conjugate with the Agilent 1290 Infinity II
LC 5991-7374EN 84
3.8 Additional Application Notes 90
3.9 More Information
Amino Acid and Cell Culture Analysis 91
4.1 Background 91
4.2 Getting Started 90
4.3 How to Guide – Amino Acid Analysis
Accurate Results with AdvanceBio End-to-End Solution
– 5991-7694EN 93
4.4 Featured Application Notes 112
4.5 Determination of Amino Acid Composition of Cell Culture Media
and Protein Hydrosylate Standard – 5991-7922EN 112
4.5 Automated Amino Acid Analysis Using an Agilent Poroshell
HPH-C18 Column – 5991-5571EN 122
4.6 More Information 130
5 Glycan Analysis 131
5.1 Background 131
5.2 Getting Started 132
5.3 How to Guide – Glycan Mapping 133
5.4 Featured Application Notes 141
5.5 Fast and Efficient HILIC Methods for Improved Analysis of Complex
Glycan Structures – 5991-4896EN 141
5.6 N-Glycan Analysis of mAbs and Other Glycoproteins with
UHPLC and Fluorescence Detection – 5991-5253EN 146
5.7 Agilent Solutions for High-throughput N-linked Glycan Profiling
from Biotherapeutics – 5991-6904EN 154
5.8 Additional Application Notes 159
5.9 More Information 159
6 Aggregate and Fragment Analysis 160
6.1 Background 160
6.2 Getting Started 161
6.3 How to Guide – Size Exclusion Chromatography for Biomolecule
Analysis – 5991-3651EN 162
6.4 Featured Application Notes 180
6.5 Size Exclusion Chromatography of Biosimilar and Innovator Insulin
5991-6872EN 180
6.6 A Comprehensive Workflow to Optimize and Execute Protein
Aggregate Studies 5991-7476EN 187
6.7 Size Exclusion Chromatography in the Presence of an Anionic
Surfactant 5991-7989EN 193
6.8 Calibrating your Agilent AdvanceBio SEC Columns
– 5991-7799EN 200
6.9 Additional Application Notes 207
6.10 More Information
Charge Variant Analysis 208
7.1 Background 208
7.2 Getting Started 209
7.3 How to Guide – Ion-Exchange Chromatography
for Biomolecule Analysis – 5991-5557EN 210
7.4 Featured Application Notes 228
7.5 Charge Heterogeneity Analysis of Rituximab Innovator and
Biosimilar mAbs – 5991-5557EN 228
7.6 Optimizing Protein Separations with Cation Exchange
Chromatography Using Agilent Buffer Advisor – 5991-0565EN 233
7.7 Faster Separations Using Agilent Weak Cation Exchange Columns
– 5990-9931EN 239
7.8 Additional Application Notes 243
8 Best Practices and Troubleshooting

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